ES2732956T3 - Processing and control of fluids - Google Patents

Abstract

A fluid control and processing system (310) for controlling the fluid flow of a fluid sample between a plurality of chambers (313), the system comprising: a housing (312) containing the plurality of chambers (313); a valve body (20) that includes a single main fluid processing region (30) continuously fluidly coupled with a single fluid displacement region (50), the fluid displacement region (50) being able to depressurize to extract the fluid in the fluid displacement region (50) and can be pressurized to expel the fluid from the fluid displacement region (50), including the valve body (20) a plurality of external ports (42, 46) , the main fluid processing region (30) being fluidly coupled with at least one external port, the fluid displacement region (50) being fluidly coupled with at least one external port (42, 46) of the body valve (20) and the valve body (20) being able to rotatably adjust with respect to the plurality of chambers (313) to place at least one external port selectively in fluid communication with the p lurality of cameras (313), wherein at least one of the plurality of cameras (313) is a processing chamber (314), including the processing chamber (314) a first port (326) and a second port (327) to selectively communicate with at least one of the external ports (42, 46) of the valve body (20), the processing chamber (314) providing an additional region of fluid processing, the processing chamber (314) containing a material fluid processing, which is an enrichment material that captures a target from the fluid sample or an exhaustion material that traps unwanted material from the fluid sample.

Description

DESCRIPTION

Processing and control of fluids

Background of the invention

The present invention generally relates to the manipulation of fluids and, more particularly, to a system and method for measuring and distributing fluids for processing and analysis.

Fluid analysis, such as clinical or environmental fluids, generally involves a series of processing steps, which may include chemical, optical, electrical, mechanical, thermal or acoustic processing of fluid samples. Whether it is incorporated into a desktop instrument, a disposable cartridge or a combination of the two, such processing typically involves complex sets of fluids and processing algorithms.

Conventional systems for processing fluid samples employ a series of chambers, each configured to subject the fluid sample to a specific processing step. As the fluid sample flows through the system sequentially from one chamber to another, the fluid sample is subjected to the processing steps according to a specific protocol. Because different protocols require different configurations, conventional systems that employ such sequential processing arrangements are not versatile or easily adaptable to different protocols.

US 5,731,212 describes a device comprising a cuvette defining at least one conduit. Reagents that must be mixed with a sample of fluids introduced into it are placed inside the ducts. A pneumatic pump extracts the fluid sample in the ducts to contact and mix the fluid sample with the various reagents. Mixing is carried out by moving the fluid sample through an obstruction in the conduit, whose obstruction causes turbulent flow in the fluid sample. A point in the cuvette is arranged below a detector used to measure a characteristic of the fluid sample.

US 4,726,237 describes a transfer value system, a method and an apparatus having a three element valve. A central element is sandwiched between the front and rear exterior elements. The specific movement of these elements is described with respect to the other elements. In an aspiration position, two sample segments are contained in series within the valve.

WO 02/18902 A1 describes fluid control and processing systems that facilitate the processing of a sample of fluids according to different protocols using the same apparatus. This document is included in the state of the art only by virtue of art. 54 (3) EPC.

Summary of the invention

The present invention provides an apparatus and method for handling fluids, by way of example, to determine the presence or absence of an analyte in a sample. In a specific embodiment, the apparatus employs a rotary valve configuration that allows fluid communication between a selectively fluid processing region with a plurality of chambers that include, for example, a sample chamber, a waste chamber, a chamber washing, a lysis chamber and a reagent or master mixing chamber. The fluid flow between the fluid processing region and the chambers is controlled by adjusting the position of the rotary valve. In this way, the measurement and distribution of fluids in the apparatus can be varied depending on the specific protocol. Unlike conventional devices, fluid flow is no longer limited to a specific protocol.

According to one aspect of the present description, a fluid control and processing system comprises a housing having a plurality of chambers and a valve body that includes a first fluid processing region continuously coupled fluidly with a region of fluid displacement. The fluid displacement region can be depressurized to extract the fluid in the fluid displacement region and can be pressurized to expel the fluid from the fluid displacement region. The valve body includes a plurality of external ports. The first fluid processing region is fluidly coupled with at least two of the external ports. The fluid displacement region is fluidly coupled with at least one of the external ports of the valve body. The valve body can be adjusted with respect to the housing to allow external ports to be selectively placed in fluid communication with the plurality of chambers. At least one of the plurality of chambers is a processing chamber that includes a first port and a second port to selectively communicate with at least one of the external ports of the valve body. The processing chamber provides an additional region of fluid processing.

In some embodiments, at least one of the fluid processing regions in the valve body or in the processing chamber contains a fluid processing material that is an enrichment material or a material of exhaustion. The fluid processing material may comprise at least one solid phase material. The solid phase material may comprise at least one of pearls, fibers, membranes, filter paper, glass wool, polymers and gels. The fluid processing material may comprise a filter and beads, or at least two types of beads. In specific embodiments, a single type of beads is used to perform at least two different functions that are selected from the group consisting of cell capture, cell lysis, analyte binding and unwanted material binding. In some embodiments, the processing chamber includes a reception area to receive a processing module containing an enrichment material or an exhaustion material. In a specific embodiment, at least one of the chambers is a reagent chamber containing dry or lyophilized reagents.

In some embodiments, the fluid processing material comprises at least one liquid phase material, such as ficol, dextran, polyethylene glycol and sucrose. The fluid processing material is contained in the fluid processing region by one or more frits. In a specific embodiment, the external ports are arranged on a generally flat external port surface of the valve body.

According to another aspect of the description, a fluid control and processing system comprises a housing having a plurality of chambers and at least one separation channel (e.g., for performing capillary electrophoresis or isoelectric focusing) and a body Valve that includes a fluid processing region continuously coupled fluidly with a fluid displacement region. The fluid displacement region can be depressurized to extract the fluid in the fluid displacement region and can be pressurized to expel the fluid from the fluid displacement region. The valve body includes at least one external port, the fluid processing region is fluidly coupled with the at least one external port and the fluid displacement region is fluidly coupled with at least one external port of the body of valve. The valve body can be adjusted with respect to the housing to allow the at least one external port to be selectively placed in fluid communication with the plurality of chambers and with the at least one separation channel.

In some embodiments, a plurality of electrodes are coupled to the housing to apply an electric field through at least a portion of the separation channel. The electrodes preferably comprise a pair of metal tubes at the two opposite ends of the separation channel. Deposits are provided at both ends of the separation channel and a deposit port is provided in one of the tanks to communicate with the at least one external port of the valve body.

Another aspect of the present invention is directed to a method for controlling the flow of fluid between a valve, a plurality of chambers and at least one separation channel, wherein the valve includes at least one external port and a fluid displacement region. continuously coupled fluidly with a fluid processing region that is fluidly coupled with the at least one external port. The method comprises adjusting the valve with respect to the plurality of chambers and the at least one separation channel to place the at least one external port selectively in fluid communication with the plurality of chambers and the at least one separation channel.

In some embodiments, an electric field is applied through at least a part of the separation channel. The method may comprise optically detecting bands of species in the separation channel.

According to another aspect of the invention, a fluid control and processing system comprises a housing having a plurality of chambers and a valve body that includes a fluid processing region continuously coupled fluidly with a region of fluid displacement. The fluid displacement region can be depressurized to extract the fluid in the fluid displacement region and can be pressurized to expel the fluid from the fluid displacement region. The valve body includes an external port. The fluid processing region is fluidly coupled with the external port. The fluid displacement region is fluidly coupled with the external port of the valve body. The valve body can be adjusted with respect to the housing to allow the external port to be selectively placed in fluid communication with the plurality of chambers.

In some embodiments, the valve body may be adjusted with respect to the housing to close the external port, so that the fluid displacement region and the fluid processing region are fluidly isolated from the chambers. At least one of the chambers and the fluid processing region may contain an enrichment material or an exhaustion material. The fluid displacement region can be depressurized by increasing the volume and can be pressurized by decreasing the volume. A fluid displacement member is disposed in the fluid displacement region and is mobile to adjust the volume of the fluid displacement region. An energy transmitting member is operatively coupled with the fluid processing region to transmit energy thereto to process the fluid contained therein.

In specific embodiments, the valve body includes a crossover channel.

The valve body can be adjusted with respect to the housing to place the crossover channel in fluid communication with an aspiration chamber and a source chamber to allow aspiration of a fluid from the source chamber through the crossover channel to the suction chamber. The body is rotatably adjustable around an axis. The at least one external port is disposed within a range of external port radii from the axis and the crossover channel is disposed within a range of crossover channel radii from the axis. The range of external port radios and the range of crossing channel radios do not overlap. The crossing channel can be a circular arc that extends over a common crossing channel radius from the axis.

In accordance with another aspect of the present description, a fluid control and processing system for controlling fluid flow between a plurality of chambers comprises a body that includes a fluid processing region continuously coupled fluidly with a region of fluid displacement.

The fluid displacement region can be depressurized to extract the fluid in the fluid displacement region and can be pressurized to expel the fluid from the fluid displacement region, including the body at least one external port. The fluid processing region is fluidly coupled with the at least one external port. The fluid displacement region is fluidly coupled with at least one external port of the valve body. The body can be rotatably adjusted and with respect to the plurality of chambers to place the at least one external port selectively in fluid communication with the plurality of chambers.

The systems of the invention are defined in claims 1 and 18. The methods of the invention are defined in claims 23 and 24. Preferred features of the systems and methods are defined in the dependent claims.

Brief description of the drawings

Fig. 1 is a perspective view of the fluid control and processing system according to an embodiment of the present description;

Fig. 2 is another perspective view of the system of Fig. 1;

Fig. 3 is an exploded view of the system of Fig. 1;

Fig. 4 is an exploded view of the system of Fig. 2;

Fig. 5 is an elevation view of a fluid control apparatus and a joint in the system of Fig. 1;

Fig. 6 is a bottom plan view of the fluid control apparatus and the seal of Fig. 5;

Fig. 7 is a top plan view of the fluid control apparatus and the seal of Fig. 5;

Fig. 8 is a cross-sectional view of the rotary fluid control apparatus of Fig. 7 along 8 8;

Fig. 9A-9LL are plan views from above and cross-sectional views illustrating a specific protocol for controlling and processing the fluid using the fluid control and processing system of Fig. 1;

Fig. 10 is an exploded perspective view of the fluid control and processing system according to another embodiment of the present description;

Fig. 11 is a cross-sectional view of a fluid control apparatus in the system of Fig. 10;

Fig. 12A-12N are plan views illustrating a specific protocol for controlling and processing the fluid using the fluid control and processing system of Fig. 10;

Fig. 13 is a cross-sectional view of a soft-walled chamber;

Fig. 14 is a cross-sectional view of a piston assembly;

Fig. 15 is a cross-sectional view of a lateral filtration chamber;

Fig. 16 is a top plan view of a fluid control and processing system that includes a processing chamber in accordance with an embodiment of the present invention;

Fig. 17 is a perspective view of the processing chamber of Fig. 16;

Fig. 18 is a partially cutaway sectional view of the fluid control and processing system of Fig. 16;

Fig. 19 is a sectional perspective view of the processing chamber of Fig. 16;

Fig. 20 is a perspective view of a retention member of the processing chamber of Fig. 16;

Fig. 21 is an elevation view of the retention member of Fig. 20;

Fig. 22 is a top plan view of the retaining member of Fig. 20;

Fig. 23 is a cross-sectional view of the retention member along 23-23 of Fig. 22; Fig. 24 is a sectional view of a fluid control and processing system that includes a separation channel according to another embodiment of the present invention;

Fig. 25 is a cross-sectional view of a fluid control apparatus in a fluid control and processing system according to another embodiment of the present invention; Y

Fig. 26A-26EE are plan views from above and cross-sectional views illustrating a specific protocol for controlling and processing fluids using the fluid control and processing system of Fig. 25.

Description of specific embodiments

Figs. 1-4 show a fluid control and processing system 10 that includes a housing 12 having a plurality of cameras 13. Fig. 1 shows the exposed cameras 13 for illustrative purposes. Normally, an upper cover will be provided to enclose the chambers 13. As best seen in Figs. 3 and 4, a fluid control device 16 and a reaction vessel 18 are connected to different parts of the housing 12. The device of fluid control in the embodiment shown is a rotary fluid control valve 16. The valve 16 includes a valve body 20 having a disk part 22 and a tubular part 24. The disk part 22 has a port surface generally flat external 23, as best seen in Fig. 3. The valve 16 can rotate with respect to the housing 12. The housing 12 includes a plurality of chamber ports 25 oriented towards the surface of the external port 23 of the part of disk 22 of the valve 16 (Fig. 4) to allow fluid communication between the chambers 13 and the valve 16. An optional seal or seal 26 is disposed between the disk part 22 and the housing 12. The disk part 22 in It also includes a filter or a stack of filters 27 and an outer cover 28 and a toothed periphery 29. The cover 28 can be a rigid sheath or a flexible film.

As best seen in Fig. 4, the disk part 22 includes a fluid processing region 30. As used herein, the term "fluid processing region" refers to a region in which a fluid is subject to processing, which includes, without limitation, chemical, optical, electrical, mechanical, thermal or acoustic processing. For example, chemical processing may include a catalyst; optical processing may include UV activation; electrical processing may include electroporation or electrophoresis or isoelectric focusing; mechanical processing may include mixing, filtering, pressurization and cell breakage; thermal processing may include heating or cooling; and acoustic processing may include the use of ultrasound. The fluid processing region may include an active member, such as filter 27, to facilitate fluid processing. Examples of active members include a fluid microchip, a solid phase material, a filter or a stack of filters, an affinity matrix, a magnetic separation matrix, a size exclusion column, a capillary tube or the like. Suitable solid phase materials include, without limitation, beads, fibers, membranes, filter paper, lysis paper impregnated with a lysis agent, glass wool, polymers or gels. In a specific embodiment, the fluid processing region is used to prepare a sample for processing, in addition, by way of example, in the reaction vessel 18.

As shown in Figs. 5-8, the outer shell 28 encloses the fluid processing region 30 and the lower end of the disc portion 22 of the valve 16. In Fig. 8, the processing region 30 includes a first fluid processing port 32 coupled to a first fluid processing channel 34 and a second fluid processing port 36 coupled to a second fluid processing channel 38. The first fluid processing channel 34 is coupled to a first external conduit 40 terminating at a first external port 42 on the surface of the external port 23, while the second fluid processing channel 38 is coupled to a second external conduit 44 terminating at a second external port 46 on the surface of the external port 23. A fluid displacement channel 48 is coupled to the first fluid processing channel 34 and the first conduit 40 near one end and to a fluid displacement region gone 50 on the other end. The first external conduit 40 serves as a common conduit to allow fluid communication between the first external port 42 and either or both of the first fluid processing channel 34 and the fluid displacement channel 48. The processing region 30 is in Continuous fluid communication with the fluid displacement region 50.

As shown in Figs. 6-8, external ports 42, 46 are angularly spaced from each other with respect to axis 52 of valve 16 at approximately 180 °. The external ports 42, 46 are radially spaced by the same distance from the axis 52. The axis 52 is perpendicular to the surface of the external port 23. In another embodiment, the angular spacing between the external ports 42, 46 may be different. The configuration of the channels in the disk part 22 may also be different in another embodiment. For example, the first fluid processing channel 34 and the first external conduit 40 may be inclined and coupled directly with the fluid displacement region 50, thereby eliminating the fluid displacement channel 48. The second fluid displacement channel 38 it can also be inclined and extend between the second fluid processing port 36 and the second external port 46 through a straight line, thereby eliminating the second external conduit 44. Additionally, more external channels and ports can be provided in the valve 16 As best seen in Fig. 3, a crossing channel or slot 56 is desirably provided on the surface of the external port 23. The slot 56 is curved and desirably spaced from the axis 52 by a constant radius. . In one embodiment, slot 56 is a circular arc that extends over a common radius from axis 52. As explained in more detail below, slot 56 is used to fill the container.

As shown in Fig. 8, the fluid displacement region 50 is disposed substantially within the tubular portion 24 of the valve 16 and partially extends into the disc portion 22. In a preferred embodiment, the displacement region of 50 fluids is a channel or pumping chamber. A fluid displacement member in the form of a piston or piston 54 is disposed movably in the pumping chamber 50. When the piston 54 moves upward, it expands the volume of the pumping chamber 50 to produce a suction to remove the fluid in the pumping chamber 50. When the piston 54 moves down, the volume of the pumping chamber 50 to drive the fluid out of the chamber 50 decreases. Alternatively, for example, pressurization and depressurization of the displacement region 50 can be carried out using a diaphragm, an external pressure or pneumatic control system or the like.

As the rotary valve 16 rotates around its axis 52 with respect to the housing 12 of Figs. 1-4, one of the external ports 42, 46 can be opened and fluidly coupled with one of the chambers 13 or reaction vessel 18 or both external ports 42, 46 may be blocked or closed. In this embodiment, at most, only one of the external ports 42, 46 is fluidly coupled with one of the chambers or reaction vessel 18. Other embodiments may be configured to allow both external ports 42, 46 to be coupled in a manner fluid with separate chambers or the reaction vessel 18. In this way, the valve 16 can rotate with respect to the housing 12 to allow the external ports 42, 46 to be selectively placed in fluid communication with a plurality of chambers that include the chambers 13 and the reaction vessel 18. Depending on which external port 42, 46 is open or closed and if the piston 54 moves up or down, the fluid flow in the valve 16 can change direction, the external ports 42, 46 can each change from being an inlet port to an outlet port and the flow of fluid can pass through the processing region 30 or bypass the process region nto 30. In a specific embodiment, the first external port 42 is the input port, so that the input side of the processing region 30 is closer to the fluid displacement region 50 than the output side of the processing region 30.

To demonstrate the fluid measurement and distribution function of valve 16, Fig. 9A-9LL illustrates the operation of valve 16 for a specific protocol. In Figs. 9A and 9AA, the first external port 42 is placed in fluid communication with a sample chamber 60 by rotating the valve 16 and the piston 54 is pulled upwards to extract a sample of fluids from the sample chamber 60 a through the first external conduit 40 and the fluid displacement channel 48 to the fluid displacement region 50, bypassing the processing region 30. For simplicity, the piston 54 is not shown in Fig. 9A-9LL. The valve 16 is then rotated to place the second external port 46 in fluid communication with a waste chamber 64, as shown in Figs. 9B and 9BB. The piston 54 is pushed down to drive the fluid sample through the fluid processing region 30 to the waste chamber 64. In a specific embodiment, the fluid processing region 30 includes a filter or a stack of filters. 27 to capture sample components (eg, cells, spores, microorganisms, viruses, proteins or the like) of the fluid sample as it passes there. An example of a stack of filters is described in United States Patent Application No. 09 / 584,327, commonly assigned, in process entitled "Device and Method for Cell Interruption", filed May 30, 2000. In alternative embodiments, other active members may be provided in the processing region 30. These first two steps of capturing sample components may be repeated as desired.

In Figs. 9C and 9CC, the valve 16 is rotated to place the first external port 42 in fluid communication with a wash chamber 66 and the piston 54 is pulled up to remove a wash liquid from the wash chamber 66 in the fluid displacement region 50, bypassing the processing region 30. The valve 16 is then rotated to place the second external port 46 in fluid communication with the waste chamber 64, as shown in Figs. 9D and 9DD . The piston 54 is pushed down to drive the washing liquid through the fluid processing region 30 to the waste chamber 64. The above washing steps can be repeated as desired. Intermediate washing is used to remove unwanted debris inside valve 16.

In Figs. 9E and 9EE, the valve 16 is rotated to place the first external port 42 in fluid communication with a lysis chamber 70 and the piston 54 is pulled upwards to extract a lysis fluid (e.g., a lysis reagent or buffer) of the lysis chamber 70 in the fluid displacement region 50, bypassing the processing region 30. The valve 16 is then rotated to place the second external port 46 in fluid communication with the fluid chamber. waste 64, as shown in Fig. 9F and 9FF. The piston 54 is pushed down to drive the lysis fluid through the fluid processing region 30 to the waste chamber 64. In Figs. 9G and 9GG, the valve 16 is rotated to close the external ports 42, 46. The piston 54 is pushed down to pressurize the remaining lysis liquid and the sample components captured in the fluid processing region 30. Additional energy can be applied to the mixture in the processing region 30. By way of For example, a sonic member 76, such as an ultrasonic horn, can be placed in contact with the outer shell 28 to transmit sonic energy in the processing region 30 to facilitate lysis of the sample components. In one embodiment, the outer shell 28 is made of a flexible film that is stretched under pressure to contact the sonic member 76 during lysis to allow transmission of sonic energy in the processing region 30.

The cover 28 in one embodiment is a flexible film of polymeric material, such as polypropylene, polyethylene, polyester or other polymers. The film can be either layered, e.g. eg, laminates or films can be homogeneous. Layered films are preferred, because they generally have better strength and structural integrity than homogeneous films. In particular, layered polypropylene films are currently preferred, because polypropylene is not an inhibitor of the polymerase chain reaction ( ^p C ^r ). Alternatively, the cover 28 may comprise other materials, such as a rigid piece of plastic. In a preferred embodiment, the cover 28 is an interface wall that is domed or includes reinforcing ribs, as shown, for example, in PCT Publication WO 00/73413 entitled "Device and method for cell interruption" or US Patent Application No. 09 / 972,221, commonly assigned, pending, entitled "Apparatus and method for rapid cell or virus disruption", filed on October 4, 2001.

In general, the energy transmission member that is operatively coupled to the processing region 30 to transmit energy thereto can be an ultrasonic, piezoelectric, magnetostrictive or electrostatic transducer. The power transmission member may also be an electromagnetic device having a wound coil, such as a voice coil motor or a solenoid device. It is currently preferred that the energy transmission member be a sonic member, such as an ultrasonic horn. Suitable speakers are commercially available from Sonics & Materials, Inc. which have an office at 53 Church Hill, Newton, Connecticut 06470-1614, USA. Alternatively, the sonic member may comprise a piezoelectric disk or any other type of ultrasonic transducer that can be coupled to the cover 28. In alternative embodiments, the energy transmission member may be a thermal element (eg, a heater) to transmit thermal energy to the processing region 30 or an electrical element to transmit electrical energy to the processing region 30. Additionally, multiple energy transmission members can be used simultaneously, e.g. eg, by simultaneously heating and sonicating the processing region to perform lysis of cells, spores, viruses or microorganisms trapped in the processing region.

In Figs. 9H and 9HH, the valve 16 is rotated to place the second external port 46 in fluid communication with a master or reagent mixing chamber 78 and the piston 54 is pushed down to elute the mixture from the region of Processing 30 to reagent chamber 78. Reagent chamber 78 typically contains reagents (eg, nucleic acid amplification reagents and probes) for mixing with the sample. Any excess mixing is dispensed in the waste chamber 64 through the second external port 46 after turning the valve 16 to place the port 46 in fluid communication with the waste chamber 64, as shown in Fig. 9I and 9II. The mixture is then mixed in reagent chamber 78 by alternation. This is accomplished by placing the fluid displacement region 50 in fluid communication with the reagent chamber 78, as shown in Figs. 9J and 9JJ and moving the piston 54 up and down. Alternating the mixture through the filter in the processing region 30, by way of example, allows larger particles trapped in the filter to move temporarily out of the way to allow passage through smaller particles. Reagent chamber 78 may contain dry or lyophilized reagents that are reconstituted when mixed with fluid.

In Figs. 9K, 9KK and 9K'K ', the valve 16 is rotated to place the first external port 42 in fluid communication with a first branch 84 coupled to the reaction vessel 18, while the second branch 86 which is coupled The reaction vessel 18 is placed in fluid communication with the crossing groove 56. The first branch 84 and the second branch 86 are arranged at different radii from the axis 52 of the valve 16, with the first branch 84 having a common radius with the first external port 42 and the second branch 86 having a common radius with the crossing slot 56. The crossing slot 56 is also in fluid communication with the reagent chamber 78 (Fig. 9K) and serves to bridge the gap between the reagent chamber 78 and the second branch 86 to provide a cross flow there between them. The external ports are arranged within a range of external port radii from the axis and the crossover slot is arranged within a range of crossover slot radii from the axis, where the range of the external port radii and the range Cross slot radii do not overlap. The placement of the crossing slot 56 in a radius other than the radius of the external ports 42, 46 is advantageous, because it prevents cross contamination of the crossing slot 56 by contaminants that may be present in the area near the surfaces between the valve 16 and housing 12 within the radius of the external ports 42, 46 as a result of the turning movement of the valve 16. Therefore, although other configurations of the crossover groove may be used, including those that overlap with the radius of the external ports 42, 46, the embodiment as shown is a preferred arrangement that isolates the crossing slot 56 from contamination of the area near the surfaces between the valve 16 and the housing 12 in the radius of the external ports 42, 46.

To fill the reaction vessel 18, the piston 54 is pulled upwards to extract the mixture in the reagent chamber 78 through the crossing groove 56 and the second branch 86 in the reaction vessel 18. In such an arrangement, the reaction vessel 18 is the aspiration chamber or is referred to as the first chamber and reagent chamber 78 is the source chamber or is referred to as the second chamber. The valve 16 is then rotated to place the second external port 46 in fluid communication with the first branch 84 and to close the first external port 42, as shown in Figs. 9L and 9LL. The piston 54 is pushed down to pressurize the mixture into the reaction vessel 18. The reaction vessel 18 can be inserted into a thermal reaction chamber to perform the amplification and / or detection of nucleic acid. The two branches 84, 86 allow filling and evacuation of the reaction chamber of the reaction vessel 18. The vessel can be connected to the housing 12 by ultrasonic welding, mechanical coupling or the like or it can be integrally formed with the housing 12, such as by molding. The use of a reaction vessel to analyze a sample of fluids is described in United States Patent Application No. 09 / 584,328, commonly assigned, in process entitled "Cartridge for proceeding with a chemical reaction", filed 30 May 2000.

To operate the valve 16 of Figs. 3-8, a motor, such as a stepper motor is normally coupled to the toothed periphery 29 of the disk part 22 to rotate the valve 16 with respect to the housing 12 to distribute The fluid with high precision. The engine can be controlled by computer according to the desired protocol. A linear motor or the like is normally used to drive piston 54 up and down with precision to provide accurate measurement and can also be computer controlled according to the desired protocol.

Fig. 10 shows another valve 100 that is rotatably coupled to a housing or block 102 of the fluid control channel. A reaction vessel 104 is detachably coupled to the housing 102. The valve 100 is a generally tubular member with a longitudinal axis 105, as shown in Fig. 11. A piston 106 is movably connected to the valve 100 to change the volume of the fluid displacement region 108 when the piston 106 moves up and down. A cover 109 is placed near the bottom of the valve 100. A fluid processing region 110 is disposed on the valve 100 and is in continuous fluid communication with the fluid displacement region 108. The valve 100 includes a pair of openings serving as a first port 111 and a second port 112, as best seen in Fig. 11. In the embodiment shown, ports 111, 112 are angularly spaced approximately 120 °, but the spacing may be different in alternative embodiments. A crossing channel or slot 114 is formed on the outer surface 116 of the valve 100 and generally extends in the longitudinal direction, as seen in Fig. 10. The two ports 111, 112 are arranged at different levels longitudinally offset between yes along the longitudinal axis 105 and the crossing groove 114 extends in the longitudinal direction of the axis 105 which bridges the two levels of the ports 111, 112.

The housing 102 has an opening 118 to receive the part of the valve 100 that has the ports 111, 112 and the slot 114. The inner surface 120 around the opening 118 is shaped to cooperate with the outer surface 116 of the valve 100. Although a gasket can be placed between the inner surface 120 and the outer surface 116, a preferred embodiment employs tapered or tapered surfaces 120, 116 that produce a sealing effect without the use of an additional gasket. The housing 102 includes a plurality of channels and ports and the valve 100 can rotate about its axis 105 to allow ports 111, 112 to be selectively placed in fluid communication with the plurality of channels in the housing 102. Depending on which port open or close and if the piston 106 moves up or down, the fluid flow in the valve 100 can change direction and the ports 111, 112 can each change from being an inlet port to a port of exit.

To demonstrate the fluid measurement and distribution function of the valve 100, Fig. 12A-12N illustrates the operation of the valve 100 for a specific protocol. As shown in Fig. 12A, the housing 102 includes a plurality of fluid channels. For convenience, the channels are labeled as follows: reagent channel 130, lysis channel 132, sample channel 134, wash channel 136, waste channel 138, first branch 140 and second branch 142. Channels 130-138 they extend from the inner surface 120 to an outer surface 144 that is generally flat and the branches 140, 142 extend from the inner surface 120 to another outer surface 146 that is also generally flat (Fig. 10). When assembled, the first port 111 and the channels 130-134 tend in a first transverse plane that is perpendicular to the longitudinal axis 105, while the second port 112, the channels 136, 138 and the two branches 140, 142 tend in a second transverse plane that is perpendicular to the longitudinal axis 105. The second transverse plane is longitudinally offset from the first transverse plane. For convenience, the second port 112, channels 136, 138 and branches 140, 142 are shaded to indicate that they are longitudinally offset from the first port 111 and channels 130-134. The crossing slot 114 extends longitudinally to bridge the deflection between the first and second transverse planes. A chamber body 150 is connected to the housing 102 (Fig. 10) and includes the reagent chamber, the lysis chamber, the sample chamber, the washing chamber and the waste chamber that are respectively fluidly coupled with channels 130-138. The first and second branches 140, 142 are fluidly coupled with the reaction vessel 104.

In Fig. 12A, the first port 111 is placed in fluid communication with the sample channel 134 and the piston 106 is pulled up to extract a sample of fluids in the fluid displacement region 108 (Fig. 11). Then, the valve 100 is rotated to place the second port 112 in fluid communication with the waste channel 138 and the piston 106 is pushed down to drive the fluid sample from the displacement region 108 through the processing region 110 and outward through the waste channel 138, as shown in Fig. 12B. These steps are usually repeated until a complete sample is processed through the processing region 110, by way of example, to capture components of the sample in a trapping member, such as a filter.

In Fig. 12C, the valve 100 is rotated to place the second port 112 in fluid communication with the wash channel 136 to aspirate a wash liquid into the processing region 110 by pulling the piston 106 upwards. The valve 100 is then rotated to place the second port 112 in fluid communication with the waste channel 138 and the piston 106 is pushed down to drive the washing liquid from the processing region 110 out through the waste channel 138. The above washing steps can be repeated as desired to remove unwanted debris inside valve 100.

For lysis, the valve 100 is rotated to place the first port 111 in fluid communication with the lysis channel 132 and the piston 106 is pulled up to extract a lysis liquid in the fluid displacement region 108, as is shown in Fig. 12E. In Fig. 12F, the valve 110 is rotated to close both ports 111, 112. The piston 106 is pushed down to push the lysis fluid in the processing region 110 and to pressurize the lysis fluid and the components of the sample captured in the fluid processing region 110. Additional energy may be applied to the mixture in the processing region 110 which includes, for example, the sonic energy transmitted in the processing region 110 by operationally coupling a sonic member with the cover 109 (Fig. 11).

In Fig. 12G, a desired predetermined amount of washing liquid is aspirated into the processing region 110 from the washing channel 136 through the second port 112 to dilute the mixture. The valve 100 is then rotated to place the first port 111 in fluid communication with the reagent channel 130 to discharge a predetermined amount of the mixture from the processing region 110 to the reagent chamber, as shown in Fig. 12H . The piston 106 moves up and down to stir and mix the mixture by alternation. The excess of the mixture is discharged through the second port 112 to the waste channel 138, as shown in Fig. 12I. Another wash is performed by extracting a washing fluid from the washing channel 136 through the second port 112 in the processing region 110 (Fig. 12J) and discharging the washing fluid from the processing region 110 through the second port 112 to waste channel 138 (Fig. 12K).

In Fig. 12L, the valve 100 is rotated to place the second port 112 in fluid communication with the first branch 140 coupled to the reaction vessel 104, while the second branch 142 that is coupled to the reaction vessel 104 is placed in fluid communication with the crossing slot 114. The second branch 142 is longitudinally offset from the reagent channel 130. In the position as shown in Fig. 12L, the crossing slot 114 extends longitudinally to bridge the deflection between the second branch 142 and reagent channel 130 to place them in fluid communication with each other. As a result, the fluid processing region 110 is in fluid communication, through the first branch 140, the reaction vessel 104, the second branch 142 and the crossing slot 114, with the reagent channel 130.

When the piston 106 is pulled up, the mixture in the reagent chamber is drawn from the reagent channel 130 through the cross slot 114 and the second branch 142 in the reaction vessel 104. The valve 100 is then rotated to place the second port 112 in fluid communication with the second branch 142 and to close the first port 111, as shown in Fig. 12M. The piston 106 is pushed down to pressurize the mixture into the reaction vessel 104. In Fig. 12N, the valve 100 is rotated to close the ports 111, 112 and isolate the reaction vessel 104. The reaction vessel 104 can Insert into a thermal reaction chamber to perform the amplification and / or detection of nucleic acid.

As illustrated in the previous embodiments, the fluid control and processing system is advantageously a fully contained system that is versatile and adaptable. The fluid displacement region is the motivating force to move the fluid in the system. By maintaining continuous fluid communication between the fluid displacement region and the fluid processing region, the motivating force to move the fluid in the system is fluidly coupled to the processing region at all times. The fluid displacement region (motivating force) also acts as a temporary storage area for the fluid that is conducted through the system. While the embodiments shown employ a mobile piston in the fluid displacement region as the motivating force, other mechanisms that include, e.g. eg, pneumatic pump mechanisms or the like that use pressure as the motivating force without a change in the volume of the fluid displacement region. The inlet or outlet side of the fluid processing region may be directed to any of the chambers to allow random access to reagents and other fluids. Complex protocols can be programmed with relative ease on a computer controller and then executed using the versatile fluid control and processing system. A large number of different protocols can be performed using a single platform.

In the embodiments shown, fluid control occurs when addressing a pair of ports on the valve to place only one port at a time selectively in fluid communication with the chambers. This is achieved by keeping the pair of ports out of phase with respect to the cameras. A crossover or bypass channel provides additional fluid control capability (eg, allowing convenient filling and emptying of the reaction vessel within the closed system). Of course, different port schemes can be used to achieve the desired fluid control in other embodiments. Moreover, while the embodiments shown include each single region of fluid processing in the valve body, additional processing regions may be located in the valve body if desired. Generally, the valve body needs (n + 1) ports per n processing regions.

The use of a single valve produces high manufacturing yields due to the presence of only one fault element. The concentration of fluid control and processing components results in a compact device (e.g., in the form of a small cartridge) and facilitates automated molding and assembly. As discussed above, the system advantageously includes the dilution and mixing capacity, the intermediate washing capacity and the positive pressurization capacity. The fluid pathways within the system are normally closed to minimize contamination and facilitate containment and control of fluids within the system. The reaction vessel is conveniently removable and replaceable and can be disposable in some embodiments.

The components of the fluid control and processing system can be made of various materials that are compatible with the fluids that are used. Examples of suitable materials include polymeric materials, such as polypropylene, polyethylene, polycarbonate, acrylic or nylon. The various cameras, channels, ports and Similar in the system can have various shapes and sizes.

The arrangements of apparatus and methods described above are merely illustrative of the applications of the principles of this description and many other embodiments and modifications can be made without departing from the scope of the invention as defined in the claims.

By way of example, Fig. 13 shows a soft wall chamber 200 that can be incorporated into the fluid control and processing system. Normally, an on-board reactive style cartridge requires a total volume of fluids of at least twice the total volume of reagents and samples combined in rigid systems. The use of soft-walled cameras can reduce the volume required. These cameras have flexible walls and can be formed normally using films and thermoforming. An added advantage of soft walls is that it is not necessary to provide ventilation if the walls are flexible enough to allow them to collapse when the chamber is emptied. In Fig. 13, a flexible side wall 202 separates a reagent chamber 204 and a waste chamber 206. Because the wastes are made up of the sample and the reagents, the volume required for the wastes is not more than the sum of the sample and reagents. Reagent chamber 204 contracts while waste chamber 206 expands and vice versa. This can be a closed system without connection to the outside. The configuration can reduce the overall size of the cartridge and can allow rapid changes in camera volumes. It can also eliminate ventilation and can reduce costs by reducing the number of platforms that should otherwise be built with hard tools. In one embodiment, at least two of the plurality of cameras in the system are separated by a flexible wall to allow the change of chamber volumes between the cameras.

Fig. 14 shows a piston assembly 210 that includes a piston rod 212 connected to a piston shaft 214 having a smaller cross section than the rod 212 for conducting small amounts of fluids. The thin piston shaft 214 can bend under an applied force if it is too long. The piston rod 212 moves along the top of the barrel or casing 216, while the piston shaft 214 moves along the bottom of the barrel 216. The movement of the piston rod 212 guides the movement of the piston shaft 214 and absorbs much of the applied force, so that very little bending force is transmitted to the thin piston shaft 214.

Fig. 15 shows a side chamber 220 that can be incorporated into the system. The side chamber 220 includes an inlet port 222 and an outlet port 224. In this example, the side chamber 220 includes a filter 226 disposed in the inlet port 222. The fluid is directed to flow through the inlet port 222. in the side chamber 220 and outwardly through the outlet port 224 for lateral filtering. This allows filtering of a sample of fluids or the like using the fluid control system of the invention. The fluid can be recirculated to achieve better filtration through the filter 226. This prefiltering is useful for removing particles before introducing the fluid into the main chambers of the system to avoid clogging. The use of a side chamber is advantageous, by way of example, to avoid contaminating the valve and the main chambers in the system.

A sample of fluids can be introduced into the housing 12 of the fluid control and processing system 10, which can be configured as a cartridge, by various mechanisms, manual or automated. For manual addition, a measured volume of material can be placed in a receiving area of the housing 12 (e.g., one of the plurality of cameras) through an input port and then a cover is placed over the port . Alternatively, the reception area can be covered by a rubber barrier or the like and the sample is injected into the reception area by pricking the barrier with a needle and injecting the sample through the needle. Alternatively, a larger amount of sample material than that required for analysis can be added to the housing 12 and the mechanisms within the housing 12 can perform the precise measurement and aliquot division of the sample necessary for the specified protocol.

It may be desirable to place certain samples, such as tissue biopsy material, soil, feces, exudates and other complex material in another device or accessory and then place the secondary device or accessory in the housing causing a mechanical action that performs a function, such as a mixture, division or extraction. For example, a piece of tissue can be placed in the lumen of a secondary device that serves as the input port cover. When the lid is pressed into the port, the tissue is forced through a mesh that slices or otherwise divides the tissue.

For the automated introduction of samples, design features of additional cartridges or housings are employed and, in many cases, sample collection functionality is imparted directly into the housing. With certain samples, such as those that pose a risk of danger to the operator or the environment, such as human retrovirus pathogens, the transfer of the sample to the housing may pose a risk. Therefore, in one embodiment, a syringe or a sorberer can be integrated into the device to provide a means to move a sample directly to the housing. Alternatively, the device may include a venous puncture needle and a tube that forms an assembly that can be used to acquire a sample. After collection, the tube and needle are removed and discarded and the housing 12 is then placed in an instrument for processing. The advantage of this approach is that the operator or the environment is not exposed to pathogens.

The port of entry can be designed taking into account the appropriate human factors depending on the nature of the intended specimen. For example, respiratory specimens can be acquired from the lower respiratory tract as cough expectorants or as smear samples or brushes from the back of the throat or nostrils. In the first case, the inlet port may be designed to allow the patient to cough directly into the housing 12 or otherwise to facilitate spitting of the specimen expectorated into the housing. For specimens of brush or smear, the specimen is placed in the port of entry where the characteristics of the port and the closure facilitate the rupture and retention of the end of the smear or brush in the area of reception of the cartridge.

In another embodiment, the housing 12 includes one or more inlet tubes or sips that can be placed in a group of samples, so that the sample material flows into the housing 12. Alternatively, a hydrophilic wick material can function to extract a shows on the device. For example, the entire cartridge can be directly submerged in the sample and a sufficient amount of sample is absorbed into the wick material and wicks into the housing 12. Then the housing is removed and can be transported to the laboratory or analyzed directly using a portable instrument In another embodiment, the tube system can be used so that one end of the tube is in direct communication with the housing to provide a fluid interface with at least one chamber and the other end is accessible to the external environment to serve as a receiver for the sample. The tube can then be placed in a sample and serve as a sorberer. Therefore, the device may include several features to collect a sample from various different sources and to move the sample to the housing 12, thereby reducing handling and inconvenience.

Fig. 16 shows a fluid control and processing system 310 that includes a housing 312 having a plurality of cameras 313 wherein one of the cameras is a processing chamber 314. The housing 312 includes a plurality of chamber ports 325 configured to communicate with a fluid control device, such as a rotary fluid control valve similar to the rotary valve 16 in the system 10 of Fig. 1-4. The valve has a fluid displacement region similar to the fluid displacement region 50 in the system 10. The chambers 313 may include the same chambers as in the embodiment of Figs. 1-4 (ie, the sample chamber 60, the waste chamber 64, the washing chamber 66, the lysis chamber 70, the reagent chamber 78 and the reaction vessel 18). The housing 312 also includes a fluid processing region or active region similar to the fluid processing region 30 of the system 10 in Figs. 1-4. In such a configuration, the ports of the chamber 325 will be oriented towards the surface of the external port of the disk part of a rotating fluid control valve.

The processing chamber 314 has a first port 326 and a second port 327. In one example, the first port 326 may be an input port for receiving fluid and the second port 327 may be an output port for discharging fluid from the chamber. of processing 314. The processing chamber 314 is normally formed or integrally constructed in the main body of the housing 312, so that the input and output ports of the processing chamber are two of the ports of the chamber. Alternatively, the processing chamber 314 can be formed as a separate member that can be inserted into the main body of the housing 312, the member having inlet and outlet ports that align with two of the chamber ports.

The processing chamber 314 may contain a processing chamber material, such as an enrichment material or medium or an exhaustion material or medium. An enrichment material captures an objective, such as an analyte, of the fluid that passes through the processing chamber 314. An exhaustion material traps or retains unwanted material from the fluid that passes through the processing chamber 314. The Enrichment or depletion material may comprise one or more solid phase materials. In general, solid phase materials may include beads, fibers, membranes, filter paper, glass wool, polymers and gels.

For example, enrichment materials may include chromatographic materials, more particularly absorbent phase materials, such as reverse phase materials, ion exchange materials or affinity chromatographic materials in which a binding member covalently binds to an insoluble matrix. For affinity chromatographic materials, the binding member may be group specific (e.g., a lectin, enzyme cofactor, Protein A and the like) or substance specific (e.g., antibody or binding fragment of the same, antigen for a particular antibody of interest, oligonucleotide and the like). The insoluble matrix to which the joint member is attached can be particles, such as porous glass or polymeric beads, strands or glass filaments, a plurality of narrow stems or capillaries and the like. For example, the insoluble matrix may include beads functionalized with antibodies to capture antigens or haptens for an immunoassay process.

Instead of coated particles or other insoluble matrices, a coated / impregnated membrane can be employed that provides selective retention of the analyte comprising the fraction of a fluid sample while allowing the rest of the sample to flow through the membrane and out of the processing chamber. Several hydrophilic, hydrophobic and ion exchange membranes have been developed for solid phase extraction.

Another example of an enrichment material is a gel medium, which can be used to provide various different screening capacities. The enrichment channel through the processing chamber 314 serves to enrich a particular analyte comprising the fraction of a liquid sample. By varying the size of the pores of the medium, using two or more gel media of different porosity and / or providing a gradient of pore size, it can be ensured that the analyte comprising the fraction of interest of the initial sample is retained in the middle of gel.

For some enrichment materials or depletion materials, it may be necessary to employ a retention mechanism to keep the particular material in the processing chamber. Frits, such as glass frits, can be used to retain the material in the processing chamber. Fig. 18-23 show two frits 330, 332 disposed within the processing chamber 314. In the embodiment shown, frits 330, 332 are held in place by a retention structure or member 336. The retention member 336 It can be configured as a processing module or an insert that can be easily snapped into place in a reception area of the processing chamber 314 and can be conveniently disposed of as desired. As shown in Fig. 17, in a specific embodiment, the processing chamber 314 includes a reception area 329 to receive a processing module containing an enrichment material or an exhaustion material. In other embodiments, the processing module may comprise a column containing a separation material or a structure containing a separation channel for capillary electrophoresis or isoelectric focusing. The processing chamber 314 has a collection area 331 for receiving fluid that has flowed through the processing module 336.

Referring to Fig. 18-23, the processing module 336 preferably includes a peak 333 that directs the fluid in the collection area 331. The processing module includes a first frit 330 that is disposed adjacent to the first port 326 and the Second frit 332 is spaced from the first frit 330 to provide a space 338 for the enrichment material or the depletion material. In one embodiment, the fluid enters the processing chamber 314 through the first port 326, passes through the first frit 330, the enrichment material or the depletion material in space 338 and the second frit 332 and then through Gravity flows to the collection area 331 of the processing chamber over the second port 327 and exits the processing chamber 314 through port 327. The space 338 serves as another fluid processing region.

In one example, a sample fluid is removed from the sample chamber by rotating the valve to place the fluid displacement region in fluid communication with the sample chamber through the first external port. This is illustrated for the system 10 of Figs. 1-4 in Figs. 9A and 9AA, which is generally the same as the system 310 of Figs. 16-23, except for the additional processing chamber 314 in the system. 310. The sample fluid circles the fluid processing region (region 30 in system 10) and enters the fluid displacement region (region 50 in system 10). The valve (valve 16 in the system 10) is rotated to place the first external port in fluid communication with the processing chamber 314. The sample fluid is conducted from the fluid displacement region in the processing chamber 314 through from inlet port 326, bypassing the fluid processing region. As the fluid flows through the processing chamber 314 containing an enrichment material through the inlet port 326, for example, the analyte comprising the sample fraction will be retained by the enrichment material, such as a Chromatographic material in the processing chamber 314. The remaining waste part of the fluid is removed out of the processing chamber 314 through the outlet port 327 and into the fluid displacement region of the valve by rotating the valve to place the first external port in fluid communication with the output port 327 of the processing chamber 314. Then, the valve is rotated to place the first external port in fluid communication with the waste chamber (chamber 64 in system 10) and the Waste fluid is conducted from the fluid displacement region in the waste chamber. An elution liquid can then flow through the enrichment material in the processing chamber 314 to release the enriched sample fraction of the enrichment material and transport it from the processing chamber 314 to another chamber or other region, such as an active region . The elution liquid can be first drawn into the fluid displacement region of the valve from another chamber and then conducted from the fluid displacement region at the inlet port 326 of the processing chamber 314 by manipulating the rotary valve . The elution liquid and the enriched sample fraction released can be extracted from the processing chamber 314 through the outlet port 327, either in the fluid displacement region through the first external port (port 42 in system 10) bypassing the fluid processing region or through the fluid processing region (region 30 in system 10) and in the fluid displacement region through the second external port (port 46 in system 10). The rotary valve can also be manipulated to transfer the fluid to other chambers or regions of the 310 system.

In another example, a depletion material is provided in the processing chamber 314 to trap or remove unwanted material from a sample fluid. The valve can be used to transfer the sample fluid from the sample chamber to the processing chamber 314, as described above. As the fluid flows through the processing chamber 314 containing an exhaustion material through the inlet port 326, unwanted materials, such as cellular waste, contaminants or amplification inhibitors, deplete of the fluid. The remaining fluid is drawn out of the processing chamber 314 through the outlet port 327 by rotating the valve to place the fluid displacement region in fluid communication with the outlet port 327. The fluid can be extracted through the second external port (port 46 in system 10) first in the fluid processing region (region 30 in system 10) and then in the fluid displacement region of the valve. Alternatively, the fluid can be extracted through the first external port (port 42 in the system 10) in the fluid displacement region by circumvailing the fluid processing region. The fluid can be subsequently conducted from the fluid displacement region in another chamber or region of the system 310 by manipulating the rotary valve.

Instead of solid phase materials, the processing chamber 314 can house liquid phase materials, such as, for example, ficol, dextran, polyethylene glycol (PEG), sucrose and the like.

By providing one or more processing chambers in the fluid processing system 310, the system 310 becomes more versatile and is capable of performing additional sample preparation steps other than those performed in the active region or the processing region in the valve body (e.g., processing region 30 in Fig. 8), to achieve multi-stage filtration, consecutive functions and the like in a single device. Moreover, the processing chamber can be fluidly coupled with an external fluid volume to facilitate large volume processing. The processing chamber can also be fluidly coupled with an external chamber containing materials that are not desirable within the main body 312 of the fluid processing system 310.

In general, the processing regions in the processing chambers (e.g., the processing chamber 314 in Fig. 16) and in the valve body (e.g., the processing region 30 in Fig. 8) can each contain enrichment materials or depletion materials. In some embodiments, each processing region may contain one or more such materials. For example, a filter (e.g., the filter or filter stack 27 in Fig. 8) or beads can be placed in a processing region to remove unwanted materials, such as cellular waste from the sample or to Get the concentration of cells. The filter or beads can be used to bind specific targets, such as particular molecules in the sample or to eliminate specific targets, such as proteins, inhibitors or the like. In some embodiments, a processing region includes a filter and other solid phase material, such as beads, fibers or wool, for molecular isolation of molecular targets or molecular removal of molecular materials. In other embodiments, a processing region may include different types of beads, such as magnetic beads, glass beads, polymer beads and the like. The beads can be used for cell capture, cell lysis, analyte binding, unwanted material binding or the like. In some embodiments, a single type of beads can be used to perform two or more of the functions of cell capture, cell lysis, analyte binding and unwanted material binding. By way of example, the cells can adhere to the beads and lysate to release their nucleic acid content and the lysate together with the released nucleic acid can be moved to a separate region or chamber for further processing, leaving the beads and their beads behind. adherent cell waste.

In another embodiment, a separation channel for performing capillary electrophoresis (CE), isoelectric focusing (IEF) or the like is provided. This can be done before or after nucleic acid amplification. The separation channel can be a separate member that is inserted into a chamber of the fluid processing system, can be formed as a microchannel in the system housing or can be constructed in one of the system's chambers.

Fig. 24 shows a separation channel or region 350 in the fluid control and processing system 354. The separation channel 350 is normally formed as a separate member that is assembled in the system 354 and, in some embodiments, can be arranged in a chamber 352. Alternatively, the separation channel 350 may be integrally formed or constructed in the system 354. The separation channel 350 may be a thin channel or a capillary coupled between at least two electrodes, which in the specific embodiment shown they include two metal tubes 356, 358. The lower end of the channel 350 is fluidly coupled to a lower reservoir 361 that is fluidly coupled to a chamber port or reservoir port 360, while the upper end of the channel 350 is fluidly coupled to a ventilated reservoir 362 provided in a support structure 366 to support the separation channel 350. The metal tubes 356, 358 serve in as electrodes for receiving electrical energy and applying an electric field to the fluid in the separation channel 350. The conductive cables in contact with the metal tubes 356, 358 can be molded in plastic and lead to respective contact areas on the external surface of the System housing 354. A voltage source can then be connected to the contact areas to apply a voltage difference between the contact areas and, therefore, between the electrodes. Alternatively, electrodes may be provided as part of an external instrument to apply the electric field and be immersed in reservoirs at the ends of the separation channel 350. The sample fluid is normally pumped by the piston 368 of the valve 370 from the displacement region of fluids 372 through one of the external ports of the valve body (e.g., external port 342) to the separation channel 350 through reservoir port 360 and reservoir 361. A sample plug is injected into the separation channel 350 and the remaining part of the sample fluid in the tank 361 can then be extracted through the port of the chamber 360 in the fluid displacement region 372 of the valve 370 by the piston 368. The tank 362 is it can be used to introduce buffer solutions, elution solvent, reagent, rinse and wash or the like into the electrophoretic flow path of the separation channel 350.

The entities in the sample cap, such as molecules, particles, cells and the like, move through a medium contained in the separation channel 350 under the influence of the applied electric field. Depending on the nature of the entities (e.g., if they carry an electric charge), as well as the surface chemistry of the electrophoretic chamber in which electrophoresis is carried out, entities can move through the medium. under the direct influence of the applied electric field or as a result of bulk fluid flow through the passageway resulting from the application of the electric field, such as an electroosmotic flow. When the sample plug is separated into bands of species in the separation channel 350, the bands are detected, by way of example, optically by a single point detector arranged at a fixed location or by a scanning detector scanning at along the length of the channel 350. To facilitate optical detection, a part of the housing can be optically transmitting or transparent. Alternatively, the detector can be inserted into the housing and placed adjacent to channel 350 (e.g., in a chamber that houses channel 350).

Normally, separation is performed after amplification, by way of example, using the method as described above in Fig. 9A-9LL. In one example, an amplified product (eg, PCR-amplified nucleic acid) is introduced as sample in reservoir 361. The separation channel 350 is prefilled with a separation material, such as a gel or buffer. A voltage is applied across the electrodes 356, 358 to inject a sample plug from the tank 361. The rest of the sample is removed after the tank 361. Next, a buffer is introduced, such as an electrolyte solution, in the tank 361. A voltage difference is applied between the electrodes 356, 358 to form an electric field that induces the flow of a sample plug through the separation channel 350 and separates the sample plug there into bands of species, which are detected using, by way of example, a single point optical detector or a scanning detector.

Fig. 25 shows the valve 416 of another system 410 having a housing with a plurality of chambers similar to the system 10 of Fig. 1-4, except that the valve 416 has only an external port 442. The valve 416 includes a valve body 420 having a disk part 422 and a tubular part 424. The disk part 422 has a generally flat outer port surface 423. The valve 416 can rotate with respect to the housing 412 of the system 410 (see Fig. 26A and 26AA). The housing 412 includes a plurality of chamber ports oriented towards the surface of the external port 423 of the disk part 422 of the valve 416 to allow fluid communication between the chambers of the housing 412 and the valve 416. The disk part 422 includes a fluid processing region 430, a first flow channel 440 that extends between the external port 442 and the fluid processing region 430 and a second flow channel 438 that extends between the fluid processing region 430 and a fluid displacement region 450 in the tubular part 424 of the valve 416. The fluid processing region 430 is in continuous fluid communication with the fluid displacement region 450. An outer shell 428 is placed over the region of fluid processing 430. The fluid processing region 430 can be used to subject a fluid flowing there through various acoustic processing, optical, thermal, electrical, mechanical or chemical.

As shown in Fig. 25, a fluid displacement member in the form of a piston or piston 454 is movably arranged in the displacement region 450 of the tubular part 424 to move up and down along the axis 452 When the piston 454 moves up, it expands the volume of the displacement region 450 to produce a suction to extract the fluid in the region 450. When the piston 454 moves down, the volume of the displacement region 450 decreases. to drive the fluid out of the region 450. As the rotary valve 416 is rotated about its axis 452 with respect to the housing 412, the external port 442 can be fluidly coupled with one of the chambers or reaction vessel in the housing 412. Depending on the action of the piston 454, the external port 442 is either an input port or an output port.

To demonstrate the fluid measurement and distribution function of valve 416, Fig. 26A-26EE illustrate the operation of valve 416 for a specific protocol. In Figs. 26A and 26AA, the external port 442 is placed in fluid communication with a sample chamber 460 by turning the valve 416 and the piston 454 is pulled upwards to extract a sample of fluids from the sample chamber 460 through of the first flow channel 440, the fluid processing region 430 and the second flow channel 438 and in the fluid displacement region 450. For simplicity, the piston 454 is not shown in Fig. 26A-26EE.

As shown in Figs. 26B and 26BB, the valve 416 is then rotated to place the external port 442 in fluid communication with a storage chamber 470 containing a lysis fluid (e.g., a lysis reagent or tampon). The piston 454 is pushed down to transfer the fluid sample from the fluid displacement region 450 to the storage chamber 470. The piston 454 is then pulled up to extract the fluid sample and the lysis fluid from the chamber. from storage 470 to the fluid displacement region 450. The lysis liquid is mixed with the sample and lyses cells or viruses in the sample. Additional energy can be applied to the processing region 430 to aid the lysis process. By way of example, a sonic member 476, such as an ultrasonic horn, can be placed in contact with the outer shell 428 to transmit ultrasonic energy in the processing region 430 to facilitate lysis of cells or viruses in the fluid sample as the fluid flows from the fluid displacement region 450 to the storage chamber 470 and / or from the storage chamber 470 back to the fluid displacement region 450. The outer cover 428 in a preferred embodiment is an interface wall It has a dome shape or includes reinforcement ribs.

In Figs. 26C and 26CC, the valve 416 is rotated to place the external port 442 in fluid communication with a reagent chamber 478 and the piston 454 is pushed down to force the lysate to flow from the fluid processing region 430 to reagent chamber 478. Reagent chamber 478 normally contains reagents (eg, PCR reagents and fluorescent probes) for mixing with the fluid sample. The fluids are then mixed in the reagent chamber 478 by alternating the mixture between the fluid displacement region 450 and the reagent chamber 478 as the piston 454 moves up and down. In Fig. 26D, 26DD and 26D'D ', the valve 416 is rotated to place the external port 442 in fluid communication with a first branch 484 coupled to the reaction vessel 418, while the second branch 486 which is coupled to the reaction vessel 418 is placed in fluid communication with the crossing groove 456. The first branch 484 and the second branch 486 are arranged at different radii from the axis 452 of the valve 416, with the first branch 484 having a common radius with the external port 442 and the second branch 486 having a common radius with the crossing slot 456. The crossing slot 456 is also in fluid communication with the reagent chamber 478 (Fig. 26D) and serves to bridge the gap between the reagent chamber 478 and the second branch 486 to provide a cross flow there between them. The external port is arranged within a range of external port radii from the axis and the crossover slot is arranged within a range of crossover slot radii from the axis, where the range of external port radii and the range of Cross slot radii do not overlap. The placement of the crossing groove 456 in a radius other than the radius of the external port 442 is advantageous, because it prevents cross contamination of the crossing groove 456 by contaminants that may be present in the area near the surfaces between the valve 416 and the housing 412 in the radius of the external port 442 as a result of the rotary movement of the valve 416.

To fill the reaction vessel 418, the piston 454 is pulled upwards to extract the mixture in the reagent chamber 478 through the crossing groove 456 and the second branch 486 in the reaction vessel 418. In such an arrangement, the reaction vessel 418 is the aspiration chamber or is referred to as the first chamber and reagent chamber 478 is the original chamber or is referred to as the second chamber. The valve 416 is then rotated to place the external port in fluid communication with the first branch 484, as shown in Fig. 26E and 26EE. The piston 454 is pushed down to pressurize the mixture into the reaction vessel 418. The reaction vessel 418 can be inserted into a thermal reaction chamber to perform amplification and / or detection of nucleic acid. The two branches 484, 486 allow the filling and evacuation of the reaction chamber of the reaction vessel 418.

The fluid control and processing system 410 of Fig. 26-26EE is modified from the system 10 of Fig. 1-9LL to provide only an external port. Similarly, the valve 100 of Fig. 10-12 can also be modified to provide only one external port by removing one of the two external ports 111, 112 and reconfiguring the fluid channels 130-138 and the branches 140, 142 between the valve 100 and the various chambers and the reaction vessel 104.

The scope of the invention, therefore, must be determined not with reference to the above description, but instead, must be determined with reference to the appended claims.

Claims (28)

1. A fluid control and processing system (310) for controlling the fluid flow of a sample of fluids between a plurality of chambers (313), the system comprising: a housing (312) containing the plurality of cameras (313); a valve body (20) that includes a single main fluid processing region (30) continuously fluidly coupled with a single fluid displacement region (50), the fluid displacement region (50) being able to depressurize to extract the fluid in the fluid displacement region (50) and can be pressurized to expel the fluid from the fluid displacement region (50), including the valve body (20) a plurality of external ports (42, 46) , the main fluid processing region (30) being fluidly coupled with at least one external port, the fluid displacement region (50) being fluidly coupled with at least one external port (42, 46) of the body valve (20) and the valve body (20) being able to rotatably adjust with respect to the plurality of chambers (313) to place at least one external port selectively in fluid communication with the p lurality of cameras (313), wherein at least one of the plurality of cameras (313) is a processing chamber (314), the processing chamber (314) including a first port (326) and a second port (327) for selectively communicating with at least one of the external ports (42, 46) of the valve body (20), the processing chamber (314) providing an additional region of fluid processing, the processing chamber (314) containing a fluid processing material, which is an enrichment material that captures a target from the fluid sample or an exhaustion material that traps unwanted material from the fluid sample. 2. The system of claim 1, wherein the main fluid processing region (30) is fluidly coupled with at least two of the external ports (42, 46) and the valve body (20) can be adjusted rotatably with respect to the housing (312) to allow the external ports (42, 46) to be selectively placed in fluid communication with the plurality of chambers (313). 3. The system of claim 1, wherein the housing further comprises: at least one separation channel and where the valve body (20) can be rotatably adjusted with respect to the housing (312) to allow selectively placing at least one external port (42, 46) in fluid communication with the plurality of chambers (313) and with the at least one separation channel. 4. The system of claim 1, wherein the fluid processing material comprises at least one solid phase material. 5. The system of claim 4, wherein the solid phase material comprises at least one of beads, fibers, membranes, filter paper, glass wool, polymers and gels. 6. The system of claim 1, wherein the fluid processing material comprises a filter and beads. 7. The system of claim 6, wherein the fluid processing material comprises at least two types of beads. 8. The system of claim 7, wherein the at least two types of beads perform at least two different functions that are selected from the group consisting of cell capture, cell lysis, analyte binding and unwanted material binding. 9. The system of claim 1, wherein the main fluid processing region (30) contains a solid phase material that performs at least two different functions selected from the group consisting of cell capture, cell lysis, analyte binding and junction of unwanted material. 10. The system of claim 9, wherein the fluid processing material comprises at least one liquid phase material. 11. The system of claim 10, wherein the liquid phase material comprises at least one of ficol, dextran, polyethylene glycol and sucrose. 12. The system of claim 9, wherein the fluid processing material is contained in the additional fluid processing region by one or more frits. 13. The system of claim 2, wherein the external ports (42, 46) are disposed on a generally flat outer port surface (23) of the valve body (20) and wherein the valve body (20) is It can rotate about an axis (52) and with respect to the plurality of chambers (313) to selectively place the external ports (42, 46) in fluid communication with the plurality of chambers (313), the axis (52) being ) perpendicular to the surface of the external port (23) and the external ports (42, 46) being spaced from the axis (52) by a common radius. 14. The system of claim 1, wherein the main fluid processing region (30) contains a type of beads that perform at least two different functions selected from the group consisting of cell capture, cell lysis, analyte binding and junction of unwanted material. 15. The system of claim 1, wherein the processing chamber (314) includes a receiving area (329) for receiving a processing module (336) containing an enrichment material that captures a target from the fluid sample or an exhaustion material that traps unwanted material from the fluid sample. 16. The system of claim 15, wherein the processing chamber further includes a collection area (331) for receiving fluid that has flowed through the processing module (336) and wherein the processing module (336 ) includes means for retaining the enrichment or depletion material in the processing module (336) and a spout (333) for directing the fluid in the collection area (331). 17. The system of any one of claims 1 to 3, wherein at least one of the chambers (313) is a reagent chamber (78) containing dry or lyophilized reagents. 18. A fluid control and processing system (354) for controlling the fluid flow of a sample of fluids between a plurality of chambers, comprising: a housing having a plurality of chambers and at least one separation channel (350); and a valve body that includes a single main fluid processing region continuously coupled fluidly with a single fluid displacement region (372), the fluid displacement region (372) being depressurized to attract the fluid to the fluid displacement region (372) and being able to pressurize to expel the fluid from the fluid displacement region (372), including the valve body at least one external port (342), the main fluid processing region being coupled fluidly with the at least one external port (342), the fluid displacement region (372) being fluidly coupled with at least one external port (342) of the valve body and the valve body being able to adjust so rotatable with respect to the housing to allow selectively placing the at least one external port (342) in fluid communication with the plurality of chambers and with the minus a separation channel (350). 19. The system (310) of claims 3 or 18, further comprising a plurality of electrodes coupled to the housing to apply an electric field through at least a portion of the separation channel (350). 20. The system of claim 19, wherein the electrodes comprise a pair of metal tubes (356, 358) at the two opposite ends of the separation channel (350). 21. The system of claims 3 or 18, further comprising reservoirs (361, 362) fluidly coupled to opposite ends of the separation channel (350) and a reservoir port (360) fluidly coupled to one of the reservoirs (361, 362) to communicate with the at least one external port (342) of the valve body. 22. The system of claim 18, wherein at least one of the chambers is a reagent chamber containing dry or lyophilized reagents. 23. A method for controlling the flow of fluid between the valve body (20) and the plurality of chambers (313) of a system (310) as described in any one of claims 1 to 17, the method comprising: rotatably adjust the valve body (20) with respect to the plurality of chambers (313) to selectively place at least one external port (42, 46) in fluid communication with the plurality of chambers (313). 24. A method of controlling the fluid flow of a sample of fluids between a valve (370), a plurality of chambers and at least one separation channel (350), including the valve (370) at least one external port (342 ) and a single fluid displacement region (372) continuously coupled fluidly with a single main fluid processing region that is fluidly coupled with the at least one external port (342), the method comprising: adjusting the valve (370) rotatably with respect to the plurality of chambers and the at least one separation channel (350) to place the at least one external port (342) selectively in fluid communication with the plurality of chambers and the channel separation (350). 25. The method of claim 24, further comprising applying an electric field through at least a portion of the separation channel (350). 26. The method of claim 24, wherein the external port (342) is placed in fluid communication with the separation channel (350) through a reservoir (361) fluidly coupled to one end of the separation channel (350), the deposit (361) having a deposit port (360) to communicate with the at least one external port (342) of the valve. 27. The method of claim 24, further comprising optically detecting bands of species in the separation channel (350). 28. A method according to claim 24, wherein the valve (370), the plurality of chambers and the at least one separation channel (350) are of a system (354) as defined in any one of the claims 18 to 22.